1887

Abstract

An expression cassette based on the highly specific tobacco etch potyvirus (TEV) nuclear inclusion (NIa) proteinase has been developed to produce multiple proteins through the translation of a single selfprocessing polypeptide. Gene constructs encoding TEV NIa, the tobacco mosaic tobamovirus (TMV) coat protein (CP) and the soybean mosaic potyvirus (SMV) CP were used to develop transgenic tobacco plants. Proper processing of the multifunctional polypeptide was demonstrated, leading to accumulation of separate proteins . Moreover, the viral genes expressed in this way were biologically active and conferred pathogen-derived protection to TMV, TEV and potato potyvirus Y (PVY). Transgenic plants were also derived from gene constructs in which the NIa cleavage site was mutated, resulting in the accumulation of the non- processed polyprotein, as predicted. Although transgenic proteins accumulated in low amounts in all the plant lines analysed, accumulation of the mutant non-processed protein form was greatly increased in plants following infection with TEV, but not TMV, apparently as a consequence of protein stabilization.

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1997-07-01
2022-05-29
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References

  1. Carrington J., Dougherty W. 1987; Small nuclear inclusion protein encoded by a plant potyvirus genome is a protease. Journal of Virology 61:2540–2548
    [Google Scholar]
  2. Carrington J., Dougherty W. 1988; A viral cleavage site cassette: identification of amino acid sequences required for tobacco etch virus polyprotein processing. Proceedings of the National Academy of Sciences USA: 853391–3395
    [Google Scholar]
  3. De Haan P., Gielen J. J., Prins M., Wijkamp I. G., Van Schepen A., Peters D., Van Grinsven M., Goldbach R. 1992; Characterization of RNA-mediated resistance to tomato spotted wilt virus in transgenic tobacco plants. Bio/Technology 10:1133–1137
    [Google Scholar]
  4. Dougherty W., Carrington J., Cary S., Parks T. 1988; Biochemical and mutational analysis of a plant virus polyprotein cleavage site. EMBO Journal 7:1281–1287
    [Google Scholar]
  5. Farinelli L., Malmoe P., Collet G. F. 1992; Heterologous encapsidation of potato virus Y strain O (PVYO) with the transgenic coat protein of PVY strain N (PVYN) in Solanum tuberosum cv. Bintje. Bio/Technology 10:1020–1025
    [Google Scholar]
  6. Fitchen J. H., Beachy R. N. 1993; Genetically engineered protection against viruses in transgenic plants. Annual Review of Microbiology 47:739–763
    [Google Scholar]
  7. Horsch R. B., Fry J. E., Hoffmann N. L., Eicholtz D., Rogers S. G., Fraley R. T. 1985; A simple and general method for transforming genes into plants. Science 227:1229–1231
    [Google Scholar]
  8. Laemmli U. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  9. Lindbo J. A., Dougherty W. G. 1992a; Pathog en-derived resistance to a potyvirus : immune and resistant phenotypes in transgenic tobacco expressing altered forms of a potyvirus coat protein nucleotide sequence. Molecular Plant-Microbe Interactions 5:144–153
    [Google Scholar]
  10. Lindbo J. A., Dougherty W. G. 1992b; Untranslatable transcripts of the tobacco etch virus coat protein gene sequence can interfere with tobacco etch virus replication in transgenic plants and protoplasts. Virology 189: 725–733
    [Google Scholar]
  11. Lindbo J. A., Silva-Rosales L., Proebsting W. M., Dougherty W. G. 1993; Induction of a highly specific antiviral state in transgenic plants : Implications for regulation of gene expression and virus resistance. Plant Cell 5:1749–1759
    [Google Scholar]
  12. Maiti I. B., Murphy J. F., Shaw J. G., Hunt A. G. 1993; Plants that express a potyvirus proteinase gene are resistant to virus infection. Proceedings of the National Academy of Sciences USA: 906110–6114
    [Google Scholar]
  13. Marcos J. F., Beachy R. N. 1994; In vitro characterization of a cassette to accumulate multiple proteins through synthesis of a selfprocessing polypeptide. Plant Molecular Biology 24:495–503
    [Google Scholar]
  14. Mueller E., Gilbert J., Davenport G., Brigneti G., Baulcombe D. 1995; Homology-dependent resistance: transgenic virus resistance in plants related to homology-dependent gene silencing. Plant Journal 7:1001–1013
    [Google Scholar]
  15. Namba K., Pattanayek R., Stubbs G. 1989; Visualization of protein-nucleic acid interactions in a virus. Refined structure of intact tobacco mosaic virus at 2·9 Å resolution by X-ray fibre diffraction. Journal of Molecular Biology 208:307–325
    [Google Scholar]
  16. Nelson R. S., Powell-Abel P., Beachy R. N. 1987; Lesions and virus accumulation in inoculated transgenic tobacco plants expressing the coat protein gene of tobacco mosaic virus. Virology 158:126–132
    [Google Scholar]
  17. Otten L. A. B. M., Schilperoort R. A. 1978; A rapid microscale method for the detection of lysopine and nopaline dehydrogenase activities. Biochimica et Biophysica Acta 257:497–500
    [Google Scholar]
  18. Pang S., Jan F., Carney K., Stout J., Tricoli D. M., Quemada H. D., Gonsalves D. 1996; Post-transcriptional transgene silencing and consequent tospovirus resistance in transgenic lettuce are affected by transgene dosage and plant development. Plant Journal 9:899–909
    [Google Scholar]
  19. Padgett H. S., Beachy R. N. 1993; Analysis of a tobacco mosaic virus strain capable of overcoming N gene-mediated resistance. Plant Cell 5:577–586
    [Google Scholar]
  20. Powell-Abel P., Nelson R. S., De B., Hoffmann N., Rogers S. G., Fraley R. T., Beachy R. N. 1986; Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232:738–743
    [Google Scholar]
  21. Reimann-Philipp U., Beachy R. N. 1993; The mechanism(s) of coat protein-mediated resistance against tobacco mosaic virus. Seminars in Virology 4:349–356
    [Google Scholar]
  22. Restrepo-Hartwig M. A., Carrington J. C. 1992; Regulation of nuclear transport of a plant potyvirus protein by autoproteolysis. Journal of Virology 66:5662–5666
    [Google Scholar]
  23. Restrepo-Hartwig M. A., Freed D. D., Carrington J. C. 1990; Nuclear transport of plant potyviral proteins. Plant Cell 2:987–998
    [Google Scholar]
  24. Riechmann J. L., Lain S., Garcia J. A. 1992; Highlights and prospects of potyvirus molecular biology. Journal of General Virology 73:1–16
    [Google Scholar]
  25. Sambrook J., Fritsch E., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Sanders P. R., Winter J. A., Barnason A. R., Rogers S. G., Fraley R. T. 1987; Comparison of cauliflower mosaic virus 35S and nopaline synthase promoters in transgenic plants. Nucleic Acids Research 15:1543–1558
    [Google Scholar]
  27. Sanford J. C., Johnston S. A. 1985; The concept of parasite-derived resistance: deriving resistance genes from the parasite’s own genome. Journal of Theoretical Biology 113:395–405
    [Google Scholar]
  28. Stark D. M., Beachy R. N. 1989; Protection against potyvirus infection in transgenic plants : evidence for broad spectrum resistance. Bio/Technology 7:1257–1262
    [Google Scholar]
  29. Tobias I., Rast A. T. B., Maat D. Z. 1982; Tobamoviruses of pepper, eggplant, and tobacco : comparative host reactions and serological relationships. Netherland Journal of Plant Pathology 88:257–268
    [Google Scholar]
  30. Van der Vlugt R. A., Ruiter R. K., Goldbach R. 1992; Evidence forsense RNA-mediated protection to PVSN in tobacco plants transformed with the viral coat protein cistron. Plant Molecular Biology 20:631–639
    [Google Scholar]
  31. Vardi E., Sela I., Edelbaum O., Livneh O., Kuznetsova L., Stram Y. 1993; Plants transformed with a cistron of a potato virus Y protease (NIa) are resistant to virus infection. Proceedings of the National Academy of Sciences USA: 907513–1517
    [Google Scholar]
  32. Verwoerd T. C., Dekker B. M. M., Hoekema A. 1989; A small scale procedure for the rapid isolation of plant RNAs. NucleicAcids Research 17:2362
    [Google Scholar]
  33. Von Bodman S. B., Domier L. L., Farrand S. K. 1995; Expression of multiple eukaryotic genes from a single promoter in Nicotiana. Bio/Technology 13:587–591
    [Google Scholar]
  34. Wilson T. M. A. 1993; Strategies to protect crop plants against viruses : pathogen-derived resistance blossoms. Proceedings of the National Academy of Sciences USA: 903134–3141
    [Google Scholar]
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